Background: Hyperbilirubinemia is a common and benign problem in neonates worldwide. It is observed during the 1st week of life in approximately 60% of term neonates and 80% of preterm neonates. Phototherapy is most widely used as therapy for unconjugated hyperbilirubinemia. Phototherapy is related to oxidative stress and lipid peroxidation. The present study is undertaken to establish the relation between anti-oxidant status and the marker of lipid peroxidation in neonatal hyperbilirubinemia before and after phototherapy. Objectives: To estimate the levels of MDA (malondialdehyde) and vit-E levels in neonatal hyperbilirubinemia before and after phototherapy. Material and Methods: A total of 30 patients were eligible for the study who met the inclusion and exclusion criteria. Blood sample was collected from neonates of preterm/full term age 1 to 10 days with hyperbilirubinemia undergoing phototherapy. Total Bilirubin, Direct Bilirubin, MDA and vit-E levels were estimated in serum by thiobarbituric acid (TBA) method and Backer and Frank’s Method respectively. Results: The present study showed increase in Total bilirubin, Direct bilirubin, MDA and decrease in vit-E levels before phototherapy when compared to control group and same subjects after phototherapy showed decrease in Total bilirubin and Direct bilirubin, MDA and a further decrease in vit-E levels. Conclusion: From these results it is concluded though phototherapy had a beneficial effect in treatment of neonatal hyperbilirubinemia, supplementation of vit-E is necessary in addition to phototherapy.